3,4-二羟基苯乙酮通过AMPK途径降低肝细胞及肝脏组织的甘油三酯含量

3,4-Dihydroxyacetophenone reduces TG content in L02 cells and hepatic tissue via AMPK pathway

目的:探讨3,4-二羟基苯乙酮(3,4-dihydroxyacetophenone,3,4-DHAP)调节肝细胞脂质代谢的机制。方法:传代培养人正常肝细胞L02,分为正常对照组、模型组、3,4-DHAP组和辛伐他汀组;药物处理8 h后收集细胞;应用试剂盒检测细胞内甘油三酯(triglyceride,TG)含量; RT-qPCR检测细胞中AMP活化蛋白激酶(AMPactivated protein kinase,AMPK)的mRNA表达水平;采用Western blot法检测细胞AMPK、磷酸化AMPK(phosphorylated AMPK,p-AMPK)、磷酸化固醇调节元件结合蛋白1c(phosphorylated sterol regulatory element binding protein 1c,p-SREBPs-1c)和磷酸化乙酰辅酶A羧化酶(phosphorylated acetyl-CoA carboxylase,p-ACC)的蛋白水平。高脂饲养新西兰大白兔,随机分为正常对照组、模型组、3,4-DHAP组和辛伐他汀组,给药处理12周后,收集肝脏标本,油红O染色;应用试剂盒分析各组TG含量;采用Western blot法检测肝脏组织中AMPK、p-AMPK、p-SREBP-1c和p-ACC的蛋白水平。结果:在细胞水平,3,4-DHAP处理后,与模型组比较,TG含量明显降低,AMPK在mRNA及蛋白水平的表达皆显著增加,AMPK磷酸化明显增加,即活性明显增强; SREBP-1c及ACC磷酸化亦显著增多。在动物实验中,经3,4-DHAP处理后,肝脏组织中的TG含量明显减少,AMPK蛋白表达显著增多、p-AMPK、p-SREBP-1c和pACC蛋白水平明显升高。结论:3,4-DHAP可能通过AMPK途径降低肝细胞及肝脏组织中的TG。

AIM:To investigate the mechanism of 3,4-dihydroxyacetophenone(3,4-DHAP)regulating lipid metabolism in human hepatic cells(L02 cells)and rabbit hepatic tissue.METHODS:L02 cells were divided into normal control group,model group,3,4-DHAP group and simvastatin group.The cells were collected after treated with drugs for 8 h.Triglyceride(TG)content in the cells was detected by TG kit.RT-qPCR was applied to detect the mRNA expression level of AMP-activated protein kinase(AMPK).The protein levels of AMPK,phosphorylated AMPK(p-AMPK),phosphorylated sterol regulatory element binding protein-1c(p-SREBP-1c)and phosphorylated acetyl-CoA carboxylase(p-ACC)were detected by Western blot.Male New Zealand white rabbits(n=32)were randomized into normal control group,model group,3,4-DHAP group and simvastatin group.The rabbits were treated with the drugs from week 2 to week 12.At the end of week 12,all rabbits were sacrificed.The liver lipids were measured by oil red O staining,and TG content was analyzed by TG kit.The protein levels of AMPK,p-AMPK,p-SREBP-1c and p-ACC in hepatic tissue were detected by Western blot.RESULTS:In L02 cells,compared with model group,TG content in 3,4-DHAP group was significantly decreased,and the expression of AMPK at mRNA and protein levels and the protein levels of p-AMPK,p-SREBP-1c and p-ACC were significantly increased.In rabbits of 3,4-DHAP group,the TG content was significantly decreased compared with model group,and the protein levels of AMPK,p-SREBP-1c and p-ACC were significantly increased.CONCLUSION:The AMPK signaling pathway may be involved in the mechanism of 3,4-DHAP to reduce TG content in L02 cells and rabbit hepatic tissue.